Fig. 3.

The 5′UTR region of the PRL mRNA controls PRL expression in response to magnesium levels. (A) PRL-2 UTR luciferase reporter constructs used to detect translation in response to magnesium levels. HeLa cells were transfected with the indicated constructs and treated with various concentrations of magnesium for 24 h and luciferase activity was measured. Values are expressed as fold change relative to 1 mM magnesium for each construct. (B) Western blot analysis of HeLa cells incubated for 24 h in the presence of the indicated magnesium concentration. Representative of three independent experiments. (C) HeLa cells were transfected with the indicated constructs, treated with varying magnesium concentrations for 24 h, and luciferase activity was measured. Plot is representative of three independent experiments. Data are means ± SD (n = 4); ****P < 0.0001 vs. control for each magnesium concentration by two-way ANOVA. (D) HeLa cells were transfected with the indicated constructs and incubated for 24 h in either the presence or absence of magnesium followed by luciferase activity measurements. Values are expressed as fold change over 1 mM magnesium for each construct. Plot is representative of two independent experiments. Data are mean ± SD (n = 4); ****P < 0.0001 vs. 1 mM magnesium by two-way ANOVA.